Human GPR158 Antibody

Monoclonal Mouse IgG1 Clone # 1027604 Catalog Number: MAB10286

DESCRIPTION Species Reactivity Human Specificity Detects human GPR158 in direct ELISAs.

Source Monoclonal Mouse IgG1 Clone # 1027604 Purification A or G purified from hybridoma culture supernatant Immunogen Human embryonic kidney cell HEK293-derived human GPR158 protein Ala24-Gln411 Accession # Q5T848 Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (-SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS.

APPLICATIONS Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website. Recommended Sample Concentration Western Blot 1 µg/mL HEK293T human embryonic kidney cell line transfected with human GPR158 Flow Cytometry 0.25 µg/106 cells HEK293 Human Cell Line Transfected with Human GPR158 and eGFP Immunohistochemistry 5-25 µg/mL Immersion fixed paraffin-embedded sections of human brain (cortex) tissue Simple Western 25 µg/mL HEK293T human embryonic kidney cell line transfected with GPR-158 and eGFP CyTOF-ready Ready to be labeled using established conjugation methods. No BSA or other carrier that could interfere with conjugation.

DATA

Western Blot Flow Cytometry Detection of Human GPR158 by Western Detection of GPR158 in HEK293 Human Cell Line Transfected Blot. Western blot shows lysates of with Human GPR158 and eGFP by Flow Cytometry. HEK293 HEK293T human embryonic kidney cell line human embryonic kidney cell line transfected with (A) human GPR158 either mock transfected or transfected with or (B) irrelevant protein, and eGFP was stained with Mouse Anti- human GPR158. PVDF membrane was Human GPR158 Monoclonal Antibody (Catalog # MAB10286) probed with 1 µg/mL of Mouse Anti-Human followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary GPR158 Monoclonal Antibody (Catalog # Antibody (Catalog # F0101B). Quadrant markers were set based on MAB10286) followed by HRP-conjugated control antibody staining (Catalog # MAB002). Staining was Anti-Mouse IgG Secondary Antibody performed using our Staining Membrane-associated Proteins (Catalog # HAF018). A specific band was protocol. detected for GPR158 at approximately 175 kDa (as indicated). GAPDH (Catalog # MAB5718) is shown as a loading control.This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Immunohistochemistry Simple Western GPR158 in Human Brain Cortex Tissue. Detection of GPR158 by Simple GPR158 was detected in immersion fixed WesternTM . Simple Western lane view shows paraffin-embedded sections of human brain lysates of HEK293T human embryonic kidney (cortex) tissue using Mouse Anti-Human cell line either mock transfected or GPR158 Monoclonal Antibody (Catalog # transfected with GPR-158 and eGFP, loaded MAB10286) at 5 µg/mL for 1 hour at room at 0.2 mg/mL. A specific band was detected temperature followed by incubation with the for GPR158 at approximately 191 kDa (as Anti-Mouse IgG VisUCyte™ HRP Polymer indicated) using 25 µg/mL of Mouse Anti- Antibody (Catalog # VC001). Before Human GPR158 Monoclonal Antibody incubation with the primary antibody, tissue (Catalog # MAB10286) . This experiment was was subjected to heat-induced epitope conducted under reducing conditions and retrieval using Antigen Retrieval Reagent- using the 12-230 kDa separation system. Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Rev. 8/10/2021 Page 1 of 2 Human GPR158 Antibody

Monoclonal Mouse IgG1 Clone # 1027604 Catalog Number: MAB10286

PREPARATION AND STORAGE Reconstitution Reconstitute at 0.5 mg/mL in sterile PBS. Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (-SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

BACKGROUND G-protein coupled receptor 158 (GPR158) is a receptor belonging to the Class C GPCR family. It lacks the extracellular Venus flytrap module characteristic of the known members of that family and instead contains two other elements that are not typical of the class: a calcium-binding EGF-like domain and a leucine repeat region (1, 2). The mature extracellular domain of human GPR158 contains 393 amino acids (aa) and shares 89% identity with both mouse and rat GPR158. GPR158 is expressed at the highest level in the brain, but also in a variety of other tissues including retina, spleen, liver and lung (3). GPR158 was originally identified in functional screens linked with biological stress and has been implicated in the effect on cognitive processes in the brain (4, 5), and glaucoma and cancer in the periphery (4, 6).

References: 1. Jingami, H. et al. (2003) Curr. Opin. Neurobiol. 13:271. 2. Bjarnadóttir, T.K et al. (2005) . 362:70. 3. Orlandi, C. et al. (2012) J. Cell Biol. 197:711. 4. Itakura, T. et al. (2019) J. Ocul. Pharmacol. Ther. 35:203. 5. Khrimian, L. et al. (2017) J. Exp. Med. 214:2859. 6. Fenner, A. (2015) Nat. Rev. Urol. 12:182.

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